AICAR Improves Outcomes of Metabolic Syndrome and Type 2 Diabetes Induced by High-Fat Diet in C57Bl 6 Male Mice PMC

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AICAR Improves Outcomes of Metabolic Syndrome and Type 2 Diabetes Induced by High-Fat Diet in C57Bl 6 Male Mice PMC

All digitized images were quantified using Un-Scan-It gel 6.1 software (Silk Scientific, Orem, UT). Specificity of primary antibodies (negative controls) was evaluated by imaging membranes with the primary antibody omitted. The following parameters (Table 13) were determined in blood serum using a SAPPHIRE 400 automatic biochemical analyzer (Tokyo Boeki LTD, Tokyo, Japan) using Randox GB reagent kits appropriate for each parameter. The animal study was reviewed and approved by the Animal Ethics Committee of Wenzhou Medical University.

AICAr, AMPK, Cancer, and Leukemia

Quantitative PCR was performed with iQ SYBR green Supermix (Bio-Rad) using the CFX96 system from Bio-Rad. In this article, we will give a brief overview of the present knowledge on AMPK-dependent and AMPK-independent effects of AICAr. Membranes were incubated in blocking solution containing commercially available antibodies overnight at 4°C [anti-phospho(Thr172)-AMPKα (40H9), 0.1 μg/ml; Anti-AMPKα (23A3), 0.1 μg/ml; Cell Signaling Technology]. Membranes were washed and incubated 1 h with the appropriate horseradish peroxidase-conjugated secondary antibodies (0.01 μg/ml; Cell Signaling Technology, Danvers, MA) and incubated in chemiluminescent substrate (West-Femto; Pierce, Grand Island, NY).

Data Availability Statement

AICAR had also no effect towards activation of three major MAPK branches by LPS, since we observed similar phosphorylation of extracellular signal-regulated kinase (ERK), p38 MAPK, and nuclear c-Jun in macrophages stimulated with LPS in the presence or absence of AICAR (Fig. 2A,B). The central goal of this study was to investigate the pharmacological activation of AMPK by AICAR as a therapeutic strategy for the treatment of PALI. Our findings demonstrated that AICAR activates AMPK, which leads to Nrf2-mediated antioxidant https://gurufer.com.br/controversy-surrounds-halotestin-steroid-course-as/ stress and inhibition of NLRP3-related inflammation, and thus improving PALI. This study indicated that AMPK exerted an essential role in the pathological processes of PALI and presented the first evidence that pharmacological activation of AMPK by AICAR ameliorates PALI, suggesting that AICAR may be a promising therapeutic agent for the treatment of PALI. The major outcome of cytosolic LPS-triggered signalling, nuclear translocation of transcriptional LPS effector RelA, remained intact in AICAR-treated cells as well.

  • In addition, the direct effect of AICAR was also assessed by treating cells with serum from NP and RUPP and adding 20 μM AICAR directly to the media.
  • However, some hepatotoxic effects were observed when the animals, on a received standard diet (STD), were treated with AICAR starting from the first day of the study.
  • Additionally, in vascular or innate immune cells, AMPK attenuates inflammatory responses by interfering with nuclear factor – κB (NFκB) and c-Jun N-terminal kinase (JNK) pathways3,4,5,6.
  • It is a member of the SREBP family, which appears to be transcription factors that regulate the expression of genes required to synthesize cholesterol, fatty acids, and triglycerides.
  • Several energy deficit states may trigger the release of AMPK, like hypoxia or hypoglycemia.

To understand the mechanism responsible for proliferation arrest in response to AICAr, we have to go back to the role of endogenous AICAR in de novo purine synthesis that has been well-known to affect cell growth much before the discovery of the role of AICAr in AMPK activation. In 2008, Narkar et al. reported that, even in sedentary mice, 4 weeks of AICAr treatment alone enhanced running endurance by 44% and induced genes linked to oxidative metabolism in muscle cells. AICAr induced fatigue-resistant type I (slow-twitch) fiber specification, and AMPK activation by AICAr was sufficient to increase running endurance without additional exercise signals [65]. In 2012, a sports doctor and nine others of the Spanish cycling team were arrested in connection with an international network supplying the synthetic AMPK activator AICAR as a “next generation superdrug” performance-enhancing drug [67].

Next, we focused on dissecting the deeper molecular mechanism by which AICAR inhibits oxidative stress and inflammation in the liver tissues of sodium taurocholate-induced SAP rats by activating AMPK phosphorylation. We performed Western blot to test the nuclear translocation of Nrf2 and the protein expression of NLRP3 as well as its downstream proteins caspase-1 and cleaved IL-1β in hepatic tissues of sodium taurocholate-induced SAP rats after treatment with AICAR. The nuclear translocation of Nrf2 was increased following sodium taurocholate treatment, whereas AICAR supplementation further promoted the nuclear accumulation of Nrf2 (Figures 4A,C). Moreover, sodium taurocholate treatment significantly increased the hepatic expression of NLRP3, caspase-1 and cleaved-IL-1β, while AICAR supplementation reversed this phenomenon (Figures 4B,C). These findings suggest that AICAR markedly alters the nuclear accumulation of Nrf2 and inhibits NLRP3 inflammasome activation in sodium taurocholate-induced PALI rats by activating AMPK phosphorylation. Thus, we speculate that Nrf2 and NLRP3 inflammasome pathway may mediate essential parts in the protective roles of AICAR against oxidative stress and inflammation in sodium taurocholate-induced PALI rats.

LPS induces an early and potent transcriptional response, which is largely dependent on the activity of NFκB and interferon response factor 3 (IRF3) transcription factors as well as the mitogen-activated protein kinase (MAPK) signalling cascade25. AICAR also lowered transcriptional activation of an anti-inflammatory cytokine IL-10 by LPS (Fig. 1C). Blocking the LPS transcriptional response in the presence of AICAR strongly inhibited secretion of IL-6 and IL-10 into the culture medium of LPS-treated macrophages (Fig. 1D). Interestingly, TNFα secretion was only partly reduced by AICAR, which can be explained by the LPS-stimulated release of already pre-formed TNFα (Fig. 1D)27. As shown in Figure 3, AICA ribotide (AICAR) or ZMP is a normal cellular intermediate in de novo purine synthesis.